title: Identification of a novel type of WRKY transcription factor binding site in elicitor-responsive cis-sequences from Arabidopsis thaliana.
abstract: Using a combination of bioinformatics and synthetic promoters, novel elicitor-responsive cis-sequences were discovered in promoters of pathogen-upregulated genes from Arabidopsis thaliana. One group of functional sequences contains the conserved core sequence GACTTTT. This core sequence and adjacent nucleotides are essential for elicitor-responsive gene expression in a parsley protoplast system. By yeast one-hybrid screening, WRKY70 was selected with a cis-sequence harbouring the core sequence GACTTTT but no known WRKY binding site (W-box). Transactivation experiments, mutation analyses, and electrophoretic mobility shift assays demonstrate that the sequence CGACTTTT is the binding site for WRKY70 in the investigated cis-sequence and is required for WRKY70-activated gene expression. Using several cis-sequences in transactivation experiments and binding studies, the CGACTTTT sequence can be extended to propose YGACTTTT as WRKY70 binding site. This binding site, designated WT-box, is enriched in promoters of genes upregulated in a WRKY70 overexpressing line. Interestingly, functional WRKY70 binding sites are present in the promoter of WRKY30, supporting recent evidence that both factors play a role in the same regulatory network.
authors.affiliation: Institut für Genetik, Technische Universität Braunschweig, Spielmannstr. 7, 38106, Braunschweig, Germany.
authors.forename: Fabian
authors.initials: F
authors.lastname: Machens
authors.forename: Marlies
authors.initials: M
authors.lastname: Becker
authors.forename: Felix
authors.initials: F
authors.lastname: Umrath
authors.forename: Reinhard
authors.initials: R
authors.lastname: Hehl
mesh_terms: D017360:Arabidopsis / Q000166:cytology / Q000235:genetics* / Q000378:metabolism; D029681:Arabidopsis Proteins / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D024202:Electrophoretic Mobility Shift Assay; D005656:Fungal Proteins / Q000235:genetics; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000235:genetics; D009154:Mutation; D059372:Nucleotide Motifs / Q000235:genetics*; D009842:Oligopeptides / Q000235:genetics / Q000378:metabolism; D010838:Phytophthora / Q000235:genetics; D011485:Protein Binding; D011523:Protoplasts / Q000378:metabolism; D020218:Response Elements / Q000235:genetics; D012689:Sequence Homology, Nucleic Acid; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism; D015533:Transcriptional Activation; D020798:Two-Hybrid System Techniques
chemical_list: D029681:Arabidopsis Proteins; D005656:Fungal Proteins; D008562:Membrane Glycoproteins; D009842:Oligopeptides; D014157:Transcription Factors; C487183:WRKY70 protein, Arabidopsis; D008156:Luciferases